Adeno-associated viral (AAV) vectors useful for transducing adipose tissue

Abstract

The present invention relates to adeno-associated viral vector useful for transducing adipose tissue. The invention also relates to polynucleotides, plasmids, vectors and methods for the production of such adeno-associated viral vector. The invention also relates to gene therapy methods useful for the treatment of a disease that requires the regulation of the expression levels of a gene.

The invention claimed is: 
     
       1. A method for treating and/or preventing a disease which requires or may benefit from the expression of a polynucleotide of interest in the adipose tissue, the method comprising administering an adeno-associated viral vector (AAV) selected from the group consisting of AAV6, AAV7, AAV8 and AAV9 comprising a recombinant viral genome wherein said recombinant viral genome comprises adeno-associated virus Inverted Terminal Repeats (ITRs) and an expression cassette, said expression cassette comprising an adipose tissue-specific transcriptional regulatory region operatively linked to a polynucleotide of interest. 
     
     
       2. The method of  claim 1  wherein the adipose tissue-specific transcriptional regulatory region comprises a promoter region selected from the group consisting of a basal aP2 promoter and a basal UCP1 promoter. 
     
     
       3. The method of  claim 2  wherein the adipose tissue-specific transcriptional regulatory region further comprises an enhancer region operatively linked to the promoter region. 
     
     
       4. The method of  claim 3  wherein the enhancer region is selected from the group consisting of an adipose-specific aP2 enhancer and an adipose-specific UCP1 enhancer. 
     
     
       5. The method of  claim 3  wherein the transcriptional regulatory region is selected from the group consisting of:
 a polynucleotide comprising an adipose-specific aP2 enhancer and a basal murine aP2 promoter; and 
 a polynucleotide comprising the adipose-specific UCP1 enhancer and a basal rat UCP1 promoter. 
 
     
     
       6. The method of  claim 1  wherein the expression cassette further comprises a post-transcriptional regulatory region. 
     
     
       7. The method of  claim 6  wherein the post-transcriptional regulatory region is a Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE). 
     
     
       8. The method of  claim 1  wherein the polynucleotide of interest encodes a protein which is selected from the group consisting of a secreted protein which acts systemically and a protein which acts upon or in a vicinity of an adipocyte. 
     
     
       9. The method of  claim 1  wherein the polynucleotide of interest encodes a protein selected from the group consisting of: hexokinase, glucokinase, alkaline phosphatase, and vascular endothelial growth factor. 
     
     
       10. The method of  claim 1  wherein the adeno-associated virus ITRs are AAV2 ITRs. 
     
     
       11. A method for treating and/or preventing a disease which requires or may benefit from the expression of a polynucleotide of interest in the adipose tissue, the method comprising administering an adeno-associated viral vector (AAV) selected from the group consisting of AAV6, AAV7, AAV8 and AAV9 comprising a recombinant viral genome wherein said recombinant viral genome comprises adeno-associated virus inverted Terminal Repeats (ITRs) and an expression cassette, said expression cassette comprising a transcriptional regulatory region operatively linked to a polynucleotide of interest, wherein said transcriptional regulatory region comprises a constitutive promoter and wherein said AAV further comprises at least one miRNA target sequence. 
     
     
       12. The method of  claim 11  wherein the at least one miRNA target sequence is mirT122a. 
     
     
       13. The method of  claim 11  wherein the at least one miRNA target sequence is mirT1. 
     
     
       14. The method of  claim 11  wherein said AAV comprises at least one copy of mirT1 and one copy of mirT122a. 
     
     
       15. The method of  claim 1 , wherein the adeno-associated viral vector (AAV) is present in a pharmaceutical composition. 
     
     
       16. The method of  claim 1  wherein the adipose tissue comprises white adipose tissue. 
     
     
       17. The method of  claim 1  wherein the adipose tissue comprises brown adipose tissue. 
     
     
       18. The method of  claim 1  wherein the adeno-associated viral vector is administered systemically or locally. 
     
     
       19. The method of  claim 11 , wherein the adeno-associated viral vector (AAV) is present in a pharmaceutical composition. 
     
     
       20. The method of  claim 11 , wherein the adeno-associated viral vector is administered systemically or locally.